Age-related mutagenesis of driver genes in the male germline

We have adapted a new ultra-sensitive sequencing technology with extremely low error-rates to identify accurately DNMs in selected candidate driver genes in sperm that can then be analyzed by our high-throughput digital PCR method. By combining these technologies, we have, for the first time, the capability to study DNMs in candidate driver genes, and investigate their expansion and increase with paternal age in the male germ line. Using our unique expertise in ultrasensitive methodologies to detect rare mutations, in this project we propose to 1) sequence exons of driver genes to discover high frequency DNMs; 2) test selection by examining the spatial expansion of the discovered DNMs in dissected testes, and 3) analyze their increase and transmission patterns in sperm of differently aged men. We envision that this project will generate a comprehensive and powerful analysis of driver mutations in the male germline, and establish a basis for understanding this type of mutagenesis and the associated risks of delayed parenthood in our society.